Alginate encapsulation of fungal microsclerotia

Abstract

Entomopathogenic fungal strains, entomopathogenic fungal compositions, entomopathogenic fungal propagules, and methods of processing to compositions stably encapsulating entomopathogenic fungal propagules.

What is claimed is: 
     
         1 . A method of preparing fungal entomopathogenic microsclerotia composition from a culture of microsclerotia comprising:
 a) mixing the microsclerotia with an alginate salt solution to form a mixture of the microsclerotia and the alginate salt solution; and   b) adding the mixture to a multivalent cation solution to form an alginate bead encapsulating the microsclerotia;
 wherein the microsclerotia in the alginate bead have enhanced stability. 
   
     
     
         2 . The method of  claim 1 , wherein the fungal entomopathogenic microsclerotia comprises at least one strain selected from the group consisting of:  Metarhizium robertsii  15013-1 , Metarhizium robertsii  23013-3, and  Metarhizium anisopliae  3213-1. 
     
     
         3 . The method of  claim 1 , further comprising collecting the microsclerotia from the culture prior to mixing with the alginate salt solution. 
     
     
         4 . The method of  claim 3 , wherein the collecting is performed by filtration or centrifugation. 
     
     
         5 . The method of  claim 1 , wherein the multivalent cation solution comprises a salt of calcium, wherein the salt of calcium further comprises carbonate, chloride, lactate, or gluconate. 
     
     
         6 . The method of  claim 1 , further comprising drying the alginate beads encapsulating the microsclerotia. 
     
     
         7 . The method of  claim 1 , further comprising breaking the alginate beads encapsulating the microsclerotia into smaller bead fragments. 
     
     
         8 . The method of  claim 1 , further comprising coating a seed with the alginate beads encapsulating the microsclerotia, or with fragments thereof. 
     
     
         9 . The method of  claim 1 , further comprising planting the alginate beads encapsulating the microsclerotia in-furrow with a seed. 
     
     
         10 . The method of  claim 1 , further comprising applying the alginate beads encapsulating the microsclerotia to the soil proximate to a plant or plant part. 
     
     
         11 . The method of  claim 1 , further comprising applying the alginate beads encapsulating the microsclerotia to the soil prior to or after planting a seed. 
     
     
         12 . The method of  claim 1 , further comprising applying the alginate beads encapsulating the microsclerotia to a plant or plant part. 
     
     
         13 . The method of  claim 1 , wherein the alginate beads encapsulating the microsclerotia increases resistance to a plant pathogen, pest or insect. 
     
     
         14 . The method of  claim 1 , wherein the microsclerotia in the alginate bead have increased stability, increased germination, or increased establishment in the soil compared to microsclerotia not encapsulated in an alginate bead. 
     
     
         15 . The method of  claim 1 , wherein the enhanced stability comprises viability of the microsclerotia for at least 2 weeks after encapsulation. 
     
     
         16 . A composition comprising at least one entomothagenic fungal strain encapsulated in an alginate bead. 
     
     
         17 . The composition of  claim 16 , wherein the entomopthogenic fungal strain is selected from the group consisting of:  Metarhizium robertsii  15013-1 , Metarhizium robertsii  23013-3, and  Metarhizium anisopliae  3213-1. 
     
     
         18 . The composition of  claim 16 , wherein the composition further comprises a biocontrol agent or plant growth-promoting agent selected from the group consisting of bacteria, a fungus, a yeast, protozoa, a virus, an entomopathogenic nematode, a botanical extract, a protein, a nucleic acid, a secondary metabolite, and an inoculant. 
     
     
         19 . The composition of  claim 16 , wherein the composition further comprises an agrochemically active compound selected from the group consisting of an insecticide, a fungicide, a bactericide, and a nematicide. 
     
     
         20 . The composition of  claim 19 , wherein the agrochemically active compound is a fungicide. 
     
     
         21 . The composition of  claim 20 , wherein the fungicide comprises a fungicide composition selected from the group consisting of azoxystrobin, thiabendazole, fludioxonil, metalaxyl, tebuconazole, prothioconazole, ipconazole, penflufen, and sedaxane. 
     
     
         22 . The composition of  claim 16 , wherein the composition further comprises a compound selected from the group consisting of a safener, a lipo-chitooligosaccharide, a benzoxazinoid, a triglucosamine lipoglycine salt, an isoflavone, and a ryanodine receptor modulator. 
     
     
         23 . The composition of  claim 16 , further comprising a plant or plant part. 
     
     
         24 . The composition of  claim 23 , wherein the plant or the plant part is genetically modified or transgenic. 
     
     
         25 . The composition of  claim 24 , wherein the genetically modified plant part is a seed. 
     
     
         26 . The composition of  claim 24 , wherein the genetically modified plant part is a leaf. 
     
     
         27 . The composition of  claim 16 , wherein the composition increases resistance of the plant or plant part to a plant pathogen, pest or insect compared to a plant or plant part without the composition. 
     
     
         28 . The composition of  claim 20 , wherein the composition retains insecticidal activity in the presence of the fungicide or is resistant to the fungicide. 
     
     
         29 . The composition of  claim 16 , wherein the composition has enhanced stability comprising viability of the microsclerotia for at least 2 weeks after encapsulation. 
     
     
         30 . A composition comprising at least one microsclerotium of at least one entomopathological fungal strain encapsulated in an alginate bead. 
     
     
         31 . The composition of  claim 30 , wherein the entomopthogenic fungal strain is selected from the group consisting of:  Metarhizium robertsii  15013-1 , Metarhizium robertsii  23013-3, and  Metarhizium anisopliae  3213-1. 
     
     
         32 . The composition of  claim 30 , wherein the composition further comprises a biocontrol agent or plant growth-promoting agent selected from the group consisting of bacteria, a fungus, a yeast, protozoa, a virus, an entomopathogenic nematode, a botanical extract, a protein, a nucleic acid, a secondary metabolite, and an inoculant. 
     
     
         33 . The composition of  claim 30 , wherein the composition further comprises an agrochemically active compound selected from the group consisting of an insecticide, a fungicide, a bactericide, and a nematicide. 
     
     
         34 . The composition of  claim 33 , wherein the agrochemically active compound is a fungicide. 
     
     
         35 . The composition of  claim 34 , wherein the fungicide comprises a fungicide composition selected from the group consisting of azoxystrobin, thiabendazole, fludioxonil, metalaxyl, tebuconazole, prothioconazole, ipconazole, penflufen, and sedaxane. 
     
     
         36 . The composition of  claim 30 , wherein the composition further comprises a compound selected from the group consisting of a safener, a lipo-chitooligosaccharide, a benzoxazinoid, a triglucosamine lipoglycine salt, an isoflavone, and a ryanodine receptor modulator. 
     
     
         37 . The composition of  claim 30 , further comprising a plant or plant part. 
     
     
         38 . The composition of  claim 37 , wherein the plant or the plant part is genetically modified or transgenic. 
     
     
         39 . The composition of  claim 38 , wherein the genetically modified plant part is a seed. 
     
     
         40 . The composition of  claim 38 , wherein the genetically modified plant part is a leaf. 
     
     
         41 . The composition of  claim 37 , wherein the composition increases resistance of the plant or plant part to a plant pathogen, pest or insect compared to a plant or plant part without the composition. 
     
     
         42 . The composition of  claim 34 , wherein the composition retains insecticidal activity in the presence of the fungicide or is resistant to the fungicide. 
     
     
         43 . The composition of  claim 30 , wherein the composition has enhanced stability comprising viability of the microsclerotia for at least 2 weeks after encapsulation.